Nature Biotechnology21, 1369 - 1377 (2003)
Published online: 31 October 2003; | doi:10.1038/nbt899
Nonlinear magic: multiphoton microscopy in the biosciences
Warren R Zipfel, Rebecca M Williams
& Watt W Webb
School of Applied and Engineering Physics, 212 Clark Hall, Cornell University, Ithaca, New York 14853, USA.
Correspondence should be addressed to Watt W Webb www2@cornell.edu
Multiphoton microscopy (MPM) has found a niche in the world of biological imaging as the best noninvasive means of fluorescence microscopy in tissue explants and living animals. Coupled with transgenic mouse models of disease and 'smart' genetically encoded fluorescent indicators, its use is now increasing exponentially. Properly applied, it is capable of measuring calcium transients 500 m deep in a mouse brain, or quantifying blood flow by imaging shadows of blood cells as they race through capillaries. With the multitude of possibilities afforded by variations of nonlinear optics and localized photochemistry, it is possible to image collagen fibrils directly within tissue through nonlinear scattering, or release caged compounds in sub-femtoliter volumes.
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m deep in a mouse brain, or quantifying blood flow by imaging shadows of blood cells as they race through capillaries. With the multitude of possibilities afforded by variations of nonlinear optics and localized photochemistry, it is possible to image collagen fibrils directly within tissue through nonlinear scattering, or release caged compounds in sub-femtoliter volumes.
